Hemostatic and wound healing turmeric-polymer composite materials

ABSTRACT

A method and composite for treating a wound. A wound is sprayed with a composition that includes a volatile organic solvent, a biocompatible polymer and turmeric powder. The solvent evaporates during the spraying. The resulting composite stops bleeding instantly, can remain on the wound for prolonged periods and adheres with the wound, even under arterial pressure. The composite serves to promote wound healing and hemostasis in bleeding wounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to, and is a non-provisional of, U.S.provisional patent application 63/241,360 (filed Sep. 7, 2021). Thecontent of this application is hereby incorporated by reference.

BACKGROUND OF THE INVENTION

Annually, the United States reports the cost for wound care to be around$31.7 billion. Hemorrhage is the number one killer on the battlefieldfor the U.S. army. In the U.S. army, 90.9% of acute mortality wasassociated with hemorrhage and most soldiers perished before reaching asurgeon. The leading cause of death for people under the age of 46 inthe United States is due to trauma and 30 to 40 percent of civiliandeaths by traumatic injury are the result of hemorrhage. As mass killingevents in the U.S. continue to rise, Hartford Consensus Joint Committeecreated a policy to increase survival in mass casualty shootings. Thepolicy views stopping hemorrhage as a crucial step to improving thesurvival of victims of an active shooter. Chronic wounds impact about 15percent (8.2 million) of Medicare beneficiaries.

Current technologies to address hemorrhage have significant limitations.These include a) making close or direct physical contact with the woundand possible exposure to patient blood, b) ingredients not appropriatefor long term use, necessitating removal of the agent from the body,once the victim is transported to a medical facility, c) use ofexpensive ingredients and lack of availability of ingredients due tosupply chain issues, d) ingredients not being biodegradable andbioabsorbable, e) agents appropriate only to arrest hemorrhage but notuseful for wound healing, f) possible side effects.

Some technologies which have been used for treating hemorrhage containzeolite, which promotes blood clotting but has side effects (thereaction of zeolites with blood is exothermic which caused second-degreeburns). Other current hemorrhage treatment agents contain kaolin—a whiteclay. The kaolin-based products are effective in arresting hemorrhage,but the ingredients are not bioresorbable. Other hemorrhage treatmentagents are not appropriate for long-term use and need to be removed fromthe body once the victim is transported to a medical facility. Thecomplications that can arise with these agents include coagulopathy, thedevelopment of progressive stages of shock. Added to these issues is thedanger associated with the release of fragments of the products into thesystemic circulation which can lead to embolus formation. Hence, theyare not appropriate for long term use. Still other hemorrhage treatmentagents are made from human blood and carry the risk of transmittinginfectious agents.

Furthermore, the world is facing several issues that make the treatmentof hemorrhage very challenging. These include a global pandemic thatoverwhelmed the hospital emergency rooms as witnessed during the COVID19 pandemic, increasing global conflicts, climate change related naturaldisasters which cause severe delays stretching into several days beforea patient can be transported to a medical facility. The issue ofaddressing hemorrhage in underdeveloped areas of the world that havelimited access to medical facilities has been a long-standing issue. Theincrease in cases of mass shootings in civilian areas where firstresponder civilians are incapable of administering the treatment due tothe fear of coming in contact with the victim's blood and the lack of areadily accessible tools to treat hemorrhage at the point of occurrenceare also other urgent matters that need to be solved immediately. Animproved technology to address at least some of these issues istherefore desired.

The discussion above is merely provided for general backgroundinformation and is not intended to be used as an aid in determining thescope of the claimed subject matter.

SUMMARY

This disclosure provides a method and composite for treating a wound. Awound is sprayed with a composition that includes a volatile organicsolvent, a biocompatible polymer and turmeric powder. The solventevaporates during the spraying. The resulting composite stops bleedinginstantly, can remain on the wound for prolonged periods and adhereswith the wound, even under arterial pressure. The composite serves topromote wound healing and hemostasis in bleeding wounds.

In a first embodiment, a method for treating a wound is provided. Themethod comprising: spraying a wound with a composition of mattercomprising: a volatile organic solvent that at least partiallyevaporates during the spraying; a biocompatible polymer; turmericpowder; and permitting the volatile organic solvent to evaporate,thereby forming a composite on the wound.

In a second embodiment, a method for forming a composite bandage isprovided. The method comprising: spraying a surface with a compositionof matter comprising: a volatile organic solvent that at least partiallyevaporates during the spraying; a biocompatible polymer; turmericpowder; permitting the volatile organic solvent to evaporate, therebyforming a composite on the surface; peeling the composite from thesurface, thereby forming the composite bandage.

In a third embodiment, a composite comprising a poly (glycolicacid-lactic acid) and a turmeric powder is provided.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1A and FIG. 1B depict two examples of spraying devices useful fordeploying the disclosed turmeric powder suspensions.

FIG. 2A depicts a mouse with a wound that is being treated with aturmeric powder mat formed in accordance with this disclosure.

FIG. 2B depicts two wounds after five days of healing wherein panel A(before) and panel B (after five days) depict a control and panels C andD show a wound before treatment with the disclosed composite (panel C)and after five days (panel D).

FIG. 3A and FIG. 3B are two scanning electron microscope (SEM) images ofa composite made according to this disclosure.

DETAILED DESCRIPTION OF THE INVENTION

This disclosure provides a composite of turmeric powder embedded in abiocompatible polymer nanofiber matrix. In one embodiment, thebiocompatible polymer is poly (lactic-co-glycolic acid), more commonlyknown as PLGA. In one embodiment, this composite is dispensed from aspraying device powered by a compressed gas. A suspension of theturmeric powder and the biocompatible polymer is formed in a volatileorganic solvent (e.g. acetone) and is loaded into the spraying device.When the spraying device is actuated, the suspension is deployed towarda target area. During deployment, the volatile organic solventevaporates mid-air to produce a composite of turmeric powder and polymernanofiber matrix that is directly applied to a wound site to arresthemorrhage and accelerate wound healing. In one embodiment, silvernanoparticles or other antibiotics are present in the suspension.

In another embodiment, bandages are produced by spraying the suspensionon a sterile surface and thereafter peeling the resulting polymernanofiber matrix off the sterile surface. The resulting bandage can bepackaged as a roll and later be directly applied to a wound. Theadhesion of the bandage to the wound is promoted by blood from the woundand can be further reinforced by wetting the bandage with an aqueoussolution such as water or commercial hand sanitizer gel.

Without wishing to be bound to any particular theory, the turmericpowder is believed to assist in hemostasis and accelerated woundhealing. The biocompatible polymer is used to produce an adherentcomposite which seals the wound and arrests hemorrhage on the spot and,in the bandage embodiment, to confer the mechanical properties andintegrity when used to patch wounds.

The resulting composite is useful for the treatment of wounds, radiationinduced tissue damage, diabetic ulcers, sealing wounds post tumorremoval for healing and elimination of resident cancerous cells at tumormargins and other skin and diseased conditions. Sealing and healing oftraumatic wounds without the need for stitches and can be used on thebattlefield, by emergency responders, ambulance staff and by the publicat all locations to prevent bleeding including situations such as publicshootings. The disclosed composite is also useful for treating splanchicand junctional wounds.

Turmeric powder is the dried powdered rhizomes of Curcuma longa of theginger family. Turmeric powder comprises 60-70% carbohydrates, 6-13%water, 6-8% protein, 5-10% fat, 3-7% dietary minerals, 3-7% essentialoils such as turmerone and germacrone, 2-7% dietary fiber, and 1-6%curcuminoids. (including curcumin). Importantly, the use of curcuminalone was not found to promote blood clotting, rather it does thereverse. It is a well-known anticoagulant (Kim D C, Ku S K, Bae J S.Anticoagulant activities of curcumin and its derivative. BMB Rep. 2012April; 45(4):221-6. doi: 10.5483/bmbrep.2012.45.4.221. PMID: 22531131).The turmeric powder may be passed through a 100 mesh (149 micron) sievesuch that the resulting particles are smaller than 149 microns indiameter. Generally, turmeric powder is present in the suspension at aconcentration between 0.1 g per mL and 2 g per ml. In one embodiment,the concentration is between 0.1 g per mL and 1 g per mL. In yet anotherembodiment, the concentration is between 0.1 g per mL and 0.5 g per mL.

Traditionally, medical fibers and sutures are prepared byelectrospinning. However, these techniques require the bioactivecompound(s) be soluble. Turmeric powder is a complex mixture of multiplecomponents, each of which has its own solubility characteristics

Some of the components of turmeric powder are water soluble, others arewater insoluble while still others are soluble but only to a limiteddegree. That makes turmeric powder an extraordinarily difficultcandidate to incorporate into a nanofiber mat or commercial hemorrhagetreatment.

A variety of biocompatible polymers may be used. Polymers include, butnot limited to, (X:Y) poly(DL-lactide-glycolide) (PLGA) where X and Ypercentage of the two monomers present in the final polymer. The valuesof X and Y can vary between 0-100% (to vary the inherent viscosity ofthe polymer between 0.1-1.75 dL per g) and include including linear andbranched polymers. Examples include poly(lactide-co-glycolide) 50:50(PLGA 50:50); poly(lactide-co-glycolide) 75:25 (PLGA 75:25);poly(lactide-co-glycolide) 25:75 (PLGA 25:75); and poly(D,L) lactide(PDLLA). In one embodiment, PLGA is used as it is biocompatible,biodegradable, bioresorbable and approved by the U.S. Food and DrugAdministration (FDA) for various applications including sutures. Otherbiocompatible polymers include poly(glycolide co-caprolactone) (PCL),polyvinyl alcohol, polydioxanone, poly(trimethylene carbonate), catgut,silk, gelatin, collagen, extracellular matrix proteins. Nylon,polyester, polyvinylidene fluoride (PVDF), polypropylene, and ultra-highmolecular weight polyethylene (UHMWPE), polytetrafluoroethylene andcyanoacrylate-based polymers.

Likewise, a variety of volatile organic solvents may be used. Examplesincludes, but are not limited to, acetone, methyl ethyl ketone, ethanol,diethyl ether and other volatile ethers, isopropanol and other shortchain alcohols, water, aqueous solutions with added salts includingbuffered solutions, acetonitrile, hexafluoroisopropanol and mixtures ofthese solvents.

Antibiotics may also be added to the formulation. Examples of suitableantibiotics include silver nanoparticles and triclosan.

Suitable spraying devices include, but are not limited to, aerosol canscommonly used to dispense spray paints, bottom and top loadingairbrushes and compressed air powder dispensers such as those used infire extinguishers. The pressurized gas used to power the airbrushinclude but are not limited to compressed air, carbon dioxide, nitrogen.In one embodiment, the spraying device uses a spraying needle with anorifice of at least 0.5 mm to prevent clogging. FIG. 1A depicts oneexample of a spraying device 100. A compressed gas cylinder 102connected to a gas regulator 104. The compressed gas cylinder 102 maybe, for example, a carbon dioxide compressed gas cylinder. Aquick-release connector 106 connects to a hose 108 which, in turn,connects to an airbrush 110 via a connector 112, such as a brass screwfitting. The airbrush 110 has a top loading container 101 that holds theturmeric powder suspension. The airbrush 110 has a top loading container101 that holds the turmeric powder suspension. FIG. 1B depicts anotherexample of a spraying device 200. Device 200 comprises a comprised gascylinder 202 connected directly to an airbrush 210. The turmeric powdersuspension is stored in bottom loading container 201. When the airbrush210 is actuated, the turmeric powder suspension is deployed through anozzle 211 of the airbrush 210. In some embodiments an extendable wand214 is present to assist in maintaining a desired distance from thetarget area. For example, the extendable wand 214 may be extended to adistance of 20 cm. The tip of the extendable wand 214 is then placedproximate the wound or other target area, thereby maintaining a distanceof 20 cm from the wound.

Exemplary Embodiment

A 20 oz. CO₂ bottle (Power tank. CYL-2140-MBK) was attached to a CO₂flow regulator from the same company (REG-4012C) and connected to aMaster Hi-Flow S622 Pro Set Dual-Action Siphon Feed Airbrush Set withNozzle (0.8 mm) and Bottle (From TCP Global Inc.) using a high pressurebraided, 5,000 PSI, hose (HSE-1241i) and coupler. The cylinder purchasedis used for paint ball, the airbrush for painting and other art relatedapplications. The regulator and connector used to assemble the portabledevice described herein were assembled based the design of the portabledevice shown in FIG. 1A.

Because the carbon dioxide cylinder was connected to the regulator withan industrial quick release connector, it afforded rapid assembly anddisassembly of the unit. The apparatus used the minimum number ofconnections as well as only a single moving part, the airbrush guntrigger. The number of connections as well the moving points werereduced in this robust design.

100 mg of pure PLGA (PLGA 50:50 ester terminated viscosity of (0.7-0.9)from Sigma or Evonik corporation or Polysciences Inc.) and 1 mL HPLCgrade acetone was combined to produce a 9% (m/v) PLGA solution. The 1 mLof PLGA solution was mixed thoroughly with 0.3 g of turmeric powder.Commercial turmeric powder sieved using a 100 mesh sieve was employedfor the studies. The CO₂ cylinder was opened to let the gas flow to theairbrush. The gas was regulated to release at a pressure 60 psi. ThePLGA and turmeric powder were placed into the solvent receptacle of theairbrush. Two types of airbrushes were tested: a siphon fed bottomloading airbrush S622 and a top loading gravity fed airbrush G 22 (bothfrom TCP Global Inc.). For the gravity feed airbrush, the PLGA andturmeric solution were placed into the barrel in 0.5 mL increments, inorder to reduce clogging. A siphon feed S622 feed airbrush with a needlesize of 0.8 mm was used. The needle diameter of 0.8 mm is much largerthan the turmeric particles overcoming any possibility of clogging andsettling of the turmeric suspension. The CO₂ pressure could be variedbetween 20-80 psi. The PLGA concentration varied between 3-9%. Theturmeric powder concentration between the range of 0.1-1 g per ml andthe distance of the target area from the tip of the airbrush from 5-50cm. In one embodiment 60 psi of CO₂ is used to power the SS622 airbrush.The mats (both control PLGA and turmeric PLGA) were directly produced ona target surface which was either a bleeding wound or a sterile surfacesuch as a glass slide when it is placed 20 cm away from the nozzle ofthe airbrush.

Due to the biocompatible nature of the composite, the composite canremain on the wound for a prolonged period of time. In one embodiment,the composite is left on the wound for a period of at least 8 hours. Inanother embodiment, the composite is left on the wound for a period ofat least 24 hours. In another embodiment, the composite is left on thewound for a period of at least 48 hours. In another embodiment, thecomposite left on the wound for a period of at least 72 hours. In stillanother embodiment the composite is left undisturbed in wounds such adeep knife wound until the wound healing is complete and the compositeis absorbed by the body.

Wound Closure Example

Referring to FIG. 2A, full thickness skin wounds of 1 cm diameter weremade in the thoracic region of CD-1 mice. A premadeturmeric-nanocomposite bandage was moistened which caused it to besecurely adhered to the wound surface. Referring to FIG. 2B, the healingof the wound was accelerated in the case of the turmeric-PLGA mat (0.3turmeric powder per mL of a 9% (m/v) solution of PLGA (50:50) inacetone) (panel C day 1 and panel D day 5) when compared to a controlPLGA mat (zero turmeric powder in a 9% (m/v) solution of PLGA (50:50) inacetone) (panel A day 1 and panel B day 5) when observed five days afterapplication. Significant contraction in the wound size can be seen byday 5 (FIG. 2B, panel D) for the turmeric power-PLGA mat treated animal.

Hemorrhage Arrest Experiment:

A mouse tail was amputated 2 cm from tip. After amputation (10 seconds)the bleeding tail was treated with turmeric powder (0.3 g per mL) in 9%(m/v) solution of PLGA (50:50) in acetone dispensed from an airbrushdevice. The turmeric composite was formed in real time on the bleedingtail and arrested hemorrhage instantly. The bleeding tail was 20 cmdistance from the airbrush nozzle which is the optimum distance at whichthe composite is formed.

Mat Example

A mat of the composite material was created from the airbrush byactuating an airbrush to dispense the suspension on a glass microscopicslide. Other suitable surfaces may also be used provided the adhesionbetween the composite and the surface is relatively low. The standardconditions were 60 PSI pressure and placing the target glass slide at 20cm from the airbrush nozzle ensures that the final mat is essentiallyfree of solvent and can be readily the peeled from the surface andformed into a roll.

FIG. 3A and FIG. 3B depict the results of scanning electron microscopy(SEM) in order to classify the morphology of the mat. The nanomat wassputter coated with a 10 nm coating of Au-PD on a glass microscopeslide. The glass microscope slide was fixed to an aluminum SEM stub. Thefibers were imaged at an accelerating voltage of 5 kilovolts and at amagnification of 100× on an AMRAY 1910 field emission scanning electronmicroscope. The resulting images show the web like formation of thenanofiber mat. SEM images of (0.3 g per mL) turmeric PLGA nanofiberswere produced by solution blow spinning using the standardizedconditions of 60 psi and mat production on a glass slide placed 20 cmfrom the nozzle of the airbrush. The images were taken at 5.0 kV and100× magnification.

Fluid Pressure

Advantageously, the composite is water impermeable and resistant to thehigh pressure the heart exerts upon the arterial walls. This ensures thecomposite is useful to stop arterial bleeding despite the fluid pressureof the escaping blood. To simulate these conditions 4 mL of 10% (m/v)PLGA/acetone solution with turmeric powder (0.3 g per mL) was sprayedonto the mouth (mouth diameter 2 cm) of a plastic bottle filled with 280mL of deionized water the distance of the airbrush nozzle from the mouthof the bottle being 20 cm. Using the mass of the water and the area ofthe bottle mouth, the exerted pressure was calculated as equal to 65mmHg which is equivalent to normal arterial pressure. Upon inversion,the water contained within the bottle did not penetrate the membrane,and the membrane itself did not rupture. This demonstrates the efficacyof turmeric-PLGA nanofibers for sealing of arteries.

Advantages:

The disclosed composition has numerous advantages. Unlike otherbandages, contact with water enhances the adhesion of the disclosedcomposite. This has been tested using the composite on mice wounds andskin. An additional case study was performed on a puncture wound on ahuman finger which was treated with the turmeric-PLGA bioactive bandagealso showed enhanced adhesion upon contact with water. Additionally, thepainful process of removing and replacing the commercial bandages, whichalso significantly disrupts wounds healing, is avoided with thedisclosed composition. Instead, the same piece initially applied willcontinue to serve as a wound healing agent over several days. Theairbrush device with turmeric-PLGA solution is ideal for coating a largesurface area such as a large glass sheet or other sheets of largesurface area (e.g. 3 foot by 3 foot). The composite maybe be directlyapplied in situations of injury that remove a large area of skin (e.g.burns) by just placing the composite on the area to be treated andwetting it with any commercial hand sanitizer.

The application method also permits a user to apply the compositewithout contacting blood which could potentially be contaminated. In oneembodiment, a distance of at least 10 cm is maintained between thespraying device and the wound during the application. In anotherembodiment, a distance of at least 20 cm is maintained.

At least one of the active ingredients in turmeric powder (curcumin) hasmoderate anti-cancer activity. Leaching studies confirmed that aconcentration of curcumin higher than 20 micro molar (higher than theIC50 of curcumin for most cancer cell lines) is released over a periodof weeks from 0.3 g per ml curcumin in 9% PLGA nanomats. The IC50 of 20micromolar for curcumin can be verified from peer reviewed literature.

This written description uses examples to disclose the invention,including the best mode, and also to enable any person skilled in theart to practice the invention, including making and using any devices orsystems and performing any incorporated methods. The patentable scope ofthe invention is defined by the claims, and may include other examplesthat occur to those skilled in the art. Such other examples are intendedto be within the scope of the claims if they have structural elementsthat do not differ from the literal language of the claims, or if theyinclude equivalent structural elements with insubstantial differencesfrom the literal language of the claims.

1. A method for treating a wound, the method comprising: spraying awound with a composition of matter comprising: a volatile organicsolvent that at least partially evaporates during the spraying; abiocompatible polymer; turmeric powder; and permitting the volatileorganic solvent to evaporate, thereby forming a composite on the wound.2. The method as recited in claim 1, further comprising leaving thecomposite on the wound for a period of at least 8 hours.
 3. The methodas recited in claim 1, wherein the biocompatible polymer is poly(glycolic acid-lactic acid).
 4. The method as recited in claim 1,wherein the composition of matter further comprises an antibiotic. 5.The method as recited in claim 1, wherein the composition of matterfurther comprises silver nanoparticles.
 6. The method as recited inclaim 1, wherein the composition of matter consists of the volatileorganic solvent, the biocompatible polymer and the turmeric powder. 7.The method as recited in claim 6, wherein the biocompatible polymer ispoly (glycolic acid-lactic acid).
 8. The method as recited in claim 7,wherein the volatile organic solvent is acetone.
 9. The method asrecited in claim 1, wherein the spraying is performed by a sprayingdevice and a distance of at least 10 cm is maintained between thespraying device and the wound during the step of spraying.
 10. Themethod as recited in claim 1, wherein the wound is an arterial woundwith arterial bleeding and the composite stops the arterial bleeding oncontact.
 11. The method as recited in claim 1, where the wound is asplanchic or junctional wound.
 12. The method as recited in claim 1,wherein the spraying occurs using a spraying device comprising anairbrush having a spraying needle with an orifice of at least 0.5 mm.13. The method as recited in claim 1, wherein the method furthercomprises applying an aqueous solution to the composite.
 14. A methodfor forming a composite bandage, the method comprising: spraying asurface with a composition of matter comprising: a volatile organicsolvent that at least partially evaporates during the spraying; abiocompatible polymer; turmeric powder; permitting the volatile organicsolvent to evaporate, thereby forming a composite on the surface;peeling the composite from the surface, thereby forming the compositebandage.
 15. The method as recited in claim 14, wherein the flat surfaceis a flat glass surface.
 16. The method as recited in claim 14, whereinthe biocompatible polymer is poly (glycolic acid-lactic acid).
 17. Themethod as recited in claim 16, wherein the composition of matterconsists of the volatile organic solvent, the biocompatible polymer andthe turmeric powder.
 18. The method as recited in claim 14, wherein thecomposition of matter further comprises silver nanoparticles.
 19. Acomposite comprising a poly (glycolic acid-lactic acid) and a turmericpowder.
 20. The composite as recited in claim 19, wherein the compositeconsists of the poly (glycolic acid-lactic acid) and the turmericpowder.